兔支气管败血波氏杆菌OMP-ELISA抗体检测方法的建立  

Development of an indirect ELISA for detecting antibodies against Bordetella bronchiseptica

作　　者：范志宇[1] 王欣[1,2] 王芳[1] 熊富强[1] 胡波[1] 徐鹏[1] 魏后军[1] 徐为中[1] 

机构地区：[1]江苏省农业科学院兽医研究所,农业部动物疫病诊断与免疫重点开放实验室,国家兽用生物制品工程技术研究中心,江苏南京210014 [2]苏州药明康德新药开发有限公司,江苏苏州215000

出　　处：《江苏农业学报》2011年第3期566-570,共5页Jiangsu Journal of Agricultural Sciences

基　　金：现代农业产业技术体系建设专项基金(nycytx-44);公益性行业(农业)科研专项(3-52肉兔);江苏省科技支撑计划--农业部分(BE2008372)

摘　　要：为简便快速检测支气管败血波氏杆菌(Bb),建立了一种特异、敏感的间接ELISA方法。通过应用超声波裂解和超速离心法提取了支气管败血波氏杆菌Ⅰ相菌外膜蛋白(OMPs),并用该蛋白包被酶标板,经特异性、敏感性和重复性试验优化后建立间接ELISA方法。结果表明,试验确定抗原的包被浓度为14.44μg/ml,可检测血清最高稀释度达1∶400;该方法有很高的特异性和敏感性,与微量凝集试验相比,敏感性提高约33倍;该方法重复性良好。In order to detect Bordetella bronchiseptica（Bb） rapidly,an indirect ELISA（enzyme-linked immuno sorbent assay） method with good specificity and sensitivity was established.In this experiment,the outer membrane proteins（OMPs） of B.bronchiseptica collected by ultrasonic and ultracentrifugation coated ELISA plates.The indirect ELISA was developed after optimization experiments of specificity,sensitivity and repeatability.The results demonstrated that optimal concentration of the antigen was 14.44 μg/ml and top detectable dilution of the serum sample was 1∶400.The ELISA method showed high specificity to Bordetella bronchiseptica（Bb） and its sensitivity was 33-fold higher than microagglutination test.

关 键 词：支气管败血波氏杆菌 外膜蛋白(OMPs) 抗体 间接ELISA 

分 类 号：S829.1[农业科学—畜牧学]